N-Linked Oligosaccharides of Aspergillus awamori Feruloyl Esterase Are Important for Thermostability and Catalysis

A unique N-linked glycosylation motif (Asn⁷⁹-Tyr-Thr) was found in the sequence of type-A feruloyl esterases from Aspergillus spp. To clarify the function of the flap, the role of N-linked oligosaccharides located in the flap region on the biochemical properties of feruloyl esterase (AwFAEA) from Aspergillus awamori expressed in Pichia pastoris was analyzed by removing the N-linked glycosylation recognition site by site-directed mutagenesis. N79 was replaced with A or Q. N-glycosylation-free N79A and N79Q mutant enzymes had lower activity than that of the glycosylated recombinant AwFAEA wild-type enzyme toward α-naphthylbutyrate (C4), α-naphthylcaprylate (C8), and phenolic acid methyl esters. Kinetic analysis of the mutant enzymes indicated that the lower catalytic efficiency was due to a combination of increased K _m and decreased k _cat for N79A, and to a considerably decreased k _cat for N79Q. N79A and N79Q mutant enzymes also exhibited considerably reduced thermostability relative to the wild-type.     

小剂量肝素治疗皮神经营养皮瓣术后静脉危象的探讨

目的：探讨小剂量肝素治疗皮神经营养皮瓣术后静脉危象的临床价值。方法：我院自2010年1月至10月共5例发生皮神经营养皮瓣术后皮瓣出现静脉危象。所有5例患者均在间断拆线的同时应用皮瓣皮下小剂量肝素注射使切缘持续缓慢渗血,并根据血运情况调整肝素使用剂量,观察皮瓣血运恢复情况及存活情况。结果：经小剂量肝素治疗后,5例皮神经营养皮瓣术后皮瓣颜色转红润,静脉危象均逐步缓解,全部皮瓣均成活,伤口无感染,愈合良好。患者对皮瓣外观和功能满意。结论：小剂量肝素皮下注射是一种简易但有效的解决皮神经营养皮瓣术后静脉危象的方法。     

足踝骨肿瘤切除后采用腓肠神经营养血管皮瓣术的修复效果及对运动功能的影响

摘要 目的：探讨足踝骨肿瘤切除后采用腓肠神经营养血管皮瓣术的修复效果及对运动功能的影响。方法：选取我院2013~2019年收治的足踝骨肿瘤切除患者83例,应用随机分组法将其分为研究组41例和对照组42例。对照组患者应用以往清创换药的保守修复方法,研究组患者应用腓肠神经营养血管皮瓣术修复方法,对比两组患者的治疗总有效率与致残率、住院费用、住院时间以及治疗前后运动功能的变化。结果：治疗后,研究组的总有效率为90.24%,显著高于对照组(64.29%,P＜0.05);两组患者住院花费比较差异无统计学意义(P＞0.05);研究组患者的住院天数上明显短于对照组(P＜0.05);研究组患者的致残率为2.44%,明显低于对照组(16.67%,P＜0.05);研究组患者修复后的Fugl-Meyer下肢运动功能评定(Fugl-Meyer assessment,FMA)和Berg平衡量表(Berg Balance Scale,BBS)评分均明显高于对照组(P＜0.05)。结论：腓肠神经营养血管皮瓣术修复术较清创换药的保守修复方法可以更有效修复足踝骨肿瘤广泛切除术后患者足踝部的创伤面,改善患者运动功能,且手术方法相对简单,安全性更高。     

背阔肌肌皮瓣修复肩背部软组织肉瘤术后缺损研究

目的：分析背阔肌肌皮瓣在肩背部软组织肉瘤扩大切除术后缺损修复中的方便性及优越性。方法：选取临床确诊肩背部软组织肉瘤患者8例,行肩背部病灶扩大切除术后,依据背阔肌肌皮瓣解剖学特点,选择合适的背阔肌肌皮瓣转移修复肩背部缺损。结果：皮瓣全部存活,随访6月至28月,肩背部外形满意,日常活动无明显影响。结论：应用背阔肌肌皮瓣修复肩背部软组织肉瘤扩大术后缺损是一种行之有效的方法。该方法简单易行,临床效果明显。     

Association between the flap endonuclease 1 gene polymorphisms and cancer susceptibility: An updated meta-analysis

Flap endonuclease 1 (FEN1) has emerged as an important enzyme in the maintenance of genomic instability and preventing carcinogenesis. The relationship between FEN1 −69G>A (rs174538)+4150G>T (rs4246215) polymorphisms and cancer susceptibility has been reported; however, results were inconclusive. In the present study, a meta-analysis of data from eligible reports was carried out to summarize the possible relationship between FEN1 polymorphisms and cancer risk. A total of 11 articles, including 20 studies with 7366 cases and 9028 controls and 18 studies with 6649 cases and 8325 controls for FEN1 rs174538 and FEN1 rs4246215 polymorphisms, respectively, were recruited for meta-analysis. Overall, meta-analyses showed that FEN1 rs174538 and rs4246215 polymorphisms are significantly associated with the decreased risk of cancer. The stratified analysis proposed that both variants were associated with protection against gastrointestinal cancer, breast cancer, hepatocellular cancer, esophageal cancer, gastric cancer, colorectal cancer, and lung cancer. In conclusion, this meta-analysis revealed an association between FEN1 polymorphisms and cancer risk. Additional studies in a larger study population that include subjects from a variety of ethnicities are warranted to further verify our findings.     

DNA repair mechanisms in dividing and non-dividing cells

DNA damage created by endogenous or exogenous genotoxic agents can exist in multiple forms, and if allowed to persist, can promote genome instability and directly lead to various human diseases, particularly cancer, neurological abnormalities, immunodeficiency and premature aging. To avoid such deleterious outcomes, cells have evolved an array of DNA repair pathways, which carry out what is typically a multiple-step process to resolve specific DNA lesions and maintain genome integrity. To fully appreciate the biological contributions of the different DNA repair systems, one must keep in mind the cellular context within which they operate. For example, the human body is composed of non-dividing and dividing cell types, including, in the brain, neurons and glial cells. We describe herein the molecular mechanisms of the different DNA repair pathways, and review their roles in non-dividing and dividing cells, with an eye toward how these pathways may regulate the development of neurological disease.     

Cryopreservation of composite tissues and transplantation: preliminary studies

Despite advances in cryobiology, the reliable cryopreservation of complex tissues has not yet been achieved. This study evaluates the viability of cryopreserved composite flaps and demonstrates the feasibility of their transplantation. Epigastric flaps were harvested from male Lewis rats. 1.5 M dimethyl sulfoxide (Me₂SO) was used as the initial cryoprotectant agent (CPA). Samples were frozen at controlled rate to −140 °C and transferred to liquid nitrogen for at least two weeks. Hematoxylin and eosin (H/E) staining, MTT tetrazolium salt assay, and factor VIII immunostaining were used to evaluate the overall histology, epithelial viability, and vascular endothelial integrity, respectively, of cryopreserved flaps. For the in vivo phase, flaps were isotransplanted to 35 recipient animals, divided into three groups: fresh (n = 10), perfused (n = 8), and cryopreserved (n = 17). Blood vessel patency was assessed via Doppler at 1, 7, and 60 days post-transplantation. For in vitro studies, cryopreserved samples (10/10) retained normal cell architecture and vascular endothelial integrity upon H/E and factor VIII staining. The viability index of cryopreserved composite flap skin (n = 10) was 11.17 ± 2.01, which was not significantly different from fresh controls (n = 10, 12.15 ± 1.32). All transplanted flaps in the fresh and perfusion groups survived with healthy color and hair growth at 60 days after operation. Survival in the cryopreserved group ranged from 2 to 60 days, with a mean of 12 days. These results demonstrate that the long term survival of cryopreserved composite tissue transplants is possible. Further studies are needed to refine protocols for the reliable cryopreservation of composite parts.     

吻合皮下静脉的带蒂皮瓣修复四肢皮肤软组织缺损的效果分析

摘要 目的：探讨与分析吻合皮下静脉的带蒂皮瓣修复四肢皮肤软组织缺损的效果。方法：选择2018年12月到2021年12月在本院创伤造成的四肢皮肤软组织缺损60例患者作为研究对象,将其随机分为吻合皮下静脉带蒂皮瓣组与传统带蒂皮瓣组各30例。吻合皮下静脉带蒂皮瓣组给予吻合皮下静脉的带蒂皮瓣修复治疗,传统带蒂皮瓣组给予常规直接覆盖创面修复治疗。结果：所有患者都顺利完成手术,吻合皮下静脉带蒂皮瓣组围手术指标时间均较传统带蒂皮瓣组少（P<0.05）。吻合皮下静脉带蒂皮瓣组术后3个月的总有效率为96.7 %,高于传统带蒂皮瓣组的76.7 %（P<0.05）。吻合皮下静脉带蒂皮瓣组术后3个月的并发症发生率较传统带蒂皮瓣组低（P<0.05）。吻合皮下静脉带蒂皮瓣组术后6个月的感觉功能恢复情况好于传统带蒂皮瓣组（P<0.05）。结论：吻合皮下静脉的带蒂皮瓣能促进患者的创面愈合,提高治疗效果,减少并发症,加快恢复患者的四肢皮肤软组织缺损。     

Delayed reconstruction of the upper digestive tract in a patient following total pharyngolaryngectomy with resection of the cervical oesophagus

Carcinoma of the hypopharynx is an uncommon disease, with an annual incidence of approximately 1 in 100,000. Post-cricoid carcinoma is more common in women and is not usually associated with tobacco and alcohol abuse. Reconstruction of large pharyngeal defects following surgery for squamous cell carcinoma is complex and often requires microvascular free tissue transfer to achieve the best oncological and functional outcomes. The most common complications of such procedures include fistulas and strictures of the neopharynx. Here, we describe a case of a female patient admitted to the Head and Neck Department at our hospital to undergo delayed reconstruction following pharyngolaryngectomy and removal of the cervical oesophagus. Several complications occurred during post-operative care, including stricture and skin dehiscence. At present, the patient is able to swallow saliva and is currently being prepared to return to a normal diet.